One strain of Pseudoalteromonas sp. E18 was isolated from the sea mud of Ningbo, Zhejiang. It can produce indigo pigment. The morphological, cultural and biochemical characteristics of the bacterium were studied. The indigo pigment was also extracted. The results showed that the maximum absorption peak of the pigment was at 579nm. The pigment was stable to UV, Na_2SO_3,It was also stable at pH 3～9. The pigment was unstable to the sunlight and high concentration H_2O_2. Temperature of 60℃～80℃ could increase the hue while temperature higher than 90℃ could reduce the hue.

OBJECTIVETo investigate whether p53 pathway participates in the effect of emodin on vascular smooth muscle cell proliferation.

METHODSThe effects of emodin on vascular smooth muscle cell proliferation were evaluated by cell count, senescent-associated beta-galactosidase staining, and annexin V staining. DNA synthesis was determined by (3)H-thymidine corporation, cell cycle was analyzed by FACS, the p53 protein level was measured by Western blot and cDNA expression array technology was used to demonstrate the effect of emodin on the simultaneous expression of a large number of genes in cultured vascular smooth muscle cells.

RESULTSEmodin at 1.6-3.1 microg/ml inhibited VSMC growth, at 6.3-12.5 microg/ml promoted VSMC aging and induced VSMC apoptosis at 25.0 microg/ml 24 hours after exposure. Unscheduled DNA synthesis, which was a sensitive indicator for DNA injury, was observed in VSMC following 24 hours emodin exposure. The mRNA and protein levels of p53 were up-regulated in a concentration-dependent manner. Proliferation/carcinogenesis-related genes were down-regulated and other genes related to cell senescence, apoptosis, and DNA damage/repair were up-regulated in VSMC after exposure to emodin for 24 hours. Emodin readily permeated VSMC membrane and mostly located in the cytoplasm and few of them in the nucleus.

CONCLUSIONSThe p53 pathway in VSMC was activated post emodin exposure in a concentration-dependent manner and which might be responsible for the observed antiproliferative effects of emodin in vascular smooth muscle cells.

Apoptosis ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; DNA Damage ; Emodin ; pharmacology ; Humans ; Myocytes, Smooth Muscle ; cytology ; drug effects ; metabolism ; RNA, Messenger ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

Adult ; Carcinoma, Squamous Cell ; diagnosis ; virology ; Cervical Intraepithelial Neoplasia ; diagnosis ; virology ; Cytological Techniques ; DNA, Viral ; analysis ; Early Detection of Cancer ; Female ; Humans ; Mass Screening ; Middle Aged ; Papillomaviridae ; genetics ; Uterine Cervical Neoplasms ; diagnosis ; virology ; Young Adult

OBJECTIVETo assess the effect of RNA interference-mediated gene silencing of plasma membrane-related Ca(2+)-ATPase-1 (PMR1) gene on the insulin secretion in islet beta cells NIT-1 in vitro.

METHODSA small interfering RNA duplex (siPMR1) corresponding to the nucleotides 337-357 of mouse PMR1 cDNA was introduced into NIT-1 cells via liposomes. The gene silencing effect was assessed by RT-PCR, and the total insulin level in the transfected cells was measured by radioimmunoassay.

RESULTSTransfection with siPMR1 resulted in obviously reduced PMR1 expression and increased insulin secretion in NIT-1 cells.

CONCLUSIONThe synthesized siPMR1 can significantly silence the expression of PMR1 and promote the secretion of insulin in the islet cells in vitro, which shed light on further studies of RNAi-based therapy of diabetes.

Animals ; Calcium-Transporting ATPases ; deficiency ; genetics ; Cell Line ; Gene Expression Regulation ; Insulin ; secretion ; Insulin-Secreting Cells ; metabolism ; secretion ; Mice ; RNA Interference ; RNA, Messenger ; genetics ; metabolism

Animals ; Fibrosis ; Genetic Testing ; Interleukin-1 ; genetics ; Kidney ; pathology ; Kidney Diseases ; complications ; diagnosis ; genetics ; Male ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Transforming Growth Factor beta ; genetics ; Ureteral Obstruction ; complications ; Vascular Cell Adhesion Molecule-1 ; genetics ; Vascular Endothelial Growth Factor A ; genetics

OBJECTIVETo clone tpn17 and tpn47 genes of Treponema pallidum and then construct their prokaryotic expression systems,to establish ELISAs based on rTpN17 and rTpN47 as antigens and to evaluate the sensitivity and specificity of the ELISAs for detection of serological diagnosis of syphilis.

METHODSThe whole length of tpn17 and tpn47 genes was amplified by PCR and then their prokaryotic expression systems were constructed. SDS-PAGE was used to measure the expression of the target recombinant proteins rTpN17 and rTpN47. Ni-NTA affinity chromatography was applied to extract rTpN17 and rTpN47, while Western blot was performed to determine the specific immunoreactivity of rTpN17 and rTpN47. By using rTpN17 and rTpN47 as the coated antigen, respectively, ELISAs (rTpN17-ELISA and rTpN47-ELISA) were established to detect serum samples from 200 healthy individuals, 25 RA patients, 17 SLE patients and 211 syphilis patients. The detection effects of the ELISAs were compared to those of TRUST and TPHA.

RESULTThe sequence similarity of the cloned tpn17 and tpn47 genes was 100 % compared with the corresponding sequences in GenBank. The expression outputs of rTpN17 and rTpN47 were approximately 37.2 % and 26.8 % of the total bacterial proteins, respectively. Both the extracted rTpN17 and rTpN47 could take place remarkable conjugation reactions to the sera with positive antibody against Treponema pallidum.The positive detection rate of TPHA (99.1%) was the highest (P<0.001). The positive detection rates of rTpN17-ELISA (85.3 %) and rTpN47-ELISA (84.3 %) were similar (P>0.05). The positive detection rates of TRUST (72.5 %) was lower than that of rTpN17-ELISA (P=0.001) but similar to that of rTpN47-ELISA (P=0.014). The detection results of all the serum samples from healthy individuals, RA patients and SLE patients were negative, whereas 7.1 % (3/42) of the samples from RA or SLE patients were positive.

CONCLUSIONrTpN17 and rTpN47 are still maintaining their original immunoreactivity. The ELISAs using rTpN17 or rTpN47 as the antigen are rapid, simple and convenient, higher sensitivity and specificity methods for serological screening and detection of syphilis.

Antibodies, Bacterial ; Antigens, Bacterial ; Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Humans ; Male ; Syphilis ; diagnosis ; Syphilis Serodiagnosis ; Treponema pallidum ; chemistry ; immunology ; isolation & purification

To study the chemical constituents of Cymbopogon citratus, isolation and purification of constituents were carried out on silica gel, Sephadex LH-20 and prepatative HPLC. The structures of the compounds were identified by physicchemical properties and spectral data analysis. Eight compounds were isolated and identified as 3beta-methoxy lanosta-9(11)-en-27-ol (1), 3beta-hydroxylanosta-9 (11)-en (2), (24S) -3beta-methoxylanosta-9(11), 25-dien-24-ol (3), 8-hydroxyl-neo-menthol (4), (2E)-3,7-dimethyl-2,7-octadiene-1, 6-diol (5), (+)-citronellol (6), 7-hydroxymenthol (7) and ethyl nonadecanoate(8). Compounds 1 is a new one. Compounds 2-3 are obtained from C. citratus for the first time.
Cymbopogon ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; Triterpenes ; chemistry

BACKGROUNDBone morphogenetic protein (BMP)-2, alkaline phosphatase (ALP), and collagen type I are known to play a critical role in the process of bone remodeling. However, the relationship between mechanical strain and the expression of BMP-2, ALP, and COL-I in osteoblasts was still unknown. The purpose of this study was to investigate the effects of different magnitudes of mechanical strain on osteoblast morphology and on the expression of BMP-2, ALP, and COL-I.

METHODSOsteoblast-like cells were flexed at four deformation rates (0, 6%, 12%, and 18% elongation). The expression of BMP-2 mRNA, ALP, and COL-I in osteoblast-like cells were determined by real-time quantitative reverse transcription polymerase chain reaction, respectively. The results were subjected to analysis of variance (ANOVA) using SPSS 13.0 statistical software.

RESULTSThe cells changed to fusiform and grew in the direction of the applied strain after the mechanical strain was loaded. Expression level of the BMP-2, ALP, and COL-I increased magnitude-dependently with mechanical loading in the experimental groups, and the 12% elongation group had the highest expression (P < 0.05).

CONCLUSIONMechanical strain can induce morphological change and a magnitude-dependent increase in the expression of BMP-2, ALP, and COL-I mRNA in osteoblast-like cells, which might influence bone remodeling in orthodontic treatment.

Alkaline Phosphatase ; metabolism ; Analysis of Variance ; Animals ; Bone Morphogenetic Protein 2 ; metabolism ; Cell Line ; Collagen ; metabolism ; Collagen Type I ; metabolism ; Mice ; Osteoblasts ; cytology ; metabolism

BACKGROUNDAnimal models are needed for the study of rapid tooth movement into the extraction socket through distraction osteogenesis of the periodontal ligament.

METHODSModified distraction devices were placed on eight dogs between the first and third mandibular premolars on the left sides; similar placement of traditional straight wise appliances on the right sides served as the control. The experimental distractors were activated (0.25 mm/d) twice a day and the control devices were activated (100 g) for two weeks with consolidation periods at weeks two, three, six, and ten. Two dogs were sacrificed at each consolidation time point; rates and patterns of tooth movement, loss of anchorage, and periapical films were evaluated, and the affected premolars and surrounding periodontal tissues were decalcified and examined histologically. General observations, X-ray periapical filming and histology examination were performed.

RESULTSDistal movement ((3.66+/-0.14) mm) measured two weeks after modified distraction exceeded that achieved using the traditional device ((1.15+/-0.21) mm; P<0.05). Loss of anchorage was minimally averaged (0.34+/-0.06) mm and (0.32+/-0.07) mm in the experimental and control sides, respectively. By radiography, apical and lateral surface root resorptions on both sides were minimal. Alveolar bone lesions were never evident. Fibroblasts were enriched in periodontal ligaments and bone spicules formed actively along directions of distraction.

CONCLUSIONSThe canine model is suitable for the study of rapid tooth movement through distraction osteogenesis of the periodontal ligament. The technique accelerates tooth movement, periodontal remodeling, alveolar bone absorption, and may induce fibroblast formation, as compared to the traditional orthodontic method, without adversely affecting root absorption, bone loss, tooth mobility and anchorage loss.

Animals ; Dogs ; Male ; Osteogenesis, Distraction ; methods ; Periodontal Ligament ; physiology ; surgery ; Root Resorption ; etiology ; Tooth Movement Techniques ; methods

BACKGROUNDThe mechanisms responsible for the occurrence of a kissing unsatisfied (KUS) result after classical crush stenting remain unclear. The present study aimed at analyzing the mechanisms and clinical significance of KUS.

METHODSTwo hundred and thirteen patients with true bifurcation lesions treated with classical crush stenting and final kissing balloon inflation (FKBI) were assigned to upper, middle, and lower groups according to the position of the side branch re-wiring assessed by visual estimation, quantitative coronary analysis (QCA) and intravascular ultrasound (IVUS). Angiographic follow-up was indexed at 12 months.

RESULTSThe upper group was characterized by a larger bifurcation angle of 55.53 degrees +/- 25.25 degrees (P = 0.030) and a longer procedural time (42.43 +/- 23.92) minutes (P = 0.015). The overall rate of KUS by visual estimation was 10.48%, with 5.4% in the upper group, 3.9% in middle group, and 36.1% in lower group (P < 0.001). For the diagnosis of KUS, visual inspection demonstrated a good correlation with both QCA and IVUS. Smaller stent diameter was the main reason for KUS in the upper group, while extra-stent side wire location, or re-wire in a low position was the main mechanism attributed to KUS in the lower group. The Lower group had more restenosis, with most restenotic lesions at a lower position of the side branch ostium. KUS (HR 1.652, 95% CI 1.332 - 2.088, P < 0.001) and re-wiring position (HR 2.341, 95% CI 1.780 - 4.329, P < 0.001) were two independent predictors of side branch restenosis. Re-wiring position (OR 0.458, 95%CI 0.336 - 0.874, P = 0.001) and side stent expansion (OR 3.122, 95%CI 2.883 - 5.061, P = 0.014) were factors predicting the findings of KUS.

CONCLUSIONSSide wire outside side stents resulted in more KUS and restenosis. Different restenotic lesion types reflected individual mechanisms contributing to the development of plaque proliferation.

Aged ; Angioplasty, Balloon, Coronary ; methods ; Coronary Disease ; therapy ; Female ; Humans ; Male ; Middle Aged ; Stents ; Treatment Outcome